The objective of this grant proposal is to provide "state of the art" methodology in protein sequencing for the UCLA scientific community. The proposals outlined by the nine (9) major users involve the use of the protein facility to aid in studies on the structure and active sites of proteins as well as tools to aid in gene cloning experiments. Much of the extensively developed recombinant DNA research work at UCLA requires protein sequencing analyses. The addition of the Milligen Sequencer will provide more sequencing time and will provide new methodologies. In particular, some projects will need to identify phospho-amino acids and other hydrophilic amino acid derivatives. The proposals listed in this grant application include studies of subcellular organelle biogenesis and function. These organelles include the myelin membrane and the unique cytosolic ribonucleoprotein, the vault. The role of lysosomal enzymes in heritable disorders is being studied using beta-hexosaminidase (Tay-Sachs) and alpha-L-iduronidase (Hurler syndrome) as models. Bacterial enzymes involved in protein secretion and membrane assembly as well as signal transducing G proteins involved in visual excitation are being studied. Structures of synthetic peptides from HIV gpl2O, gp4l, the envelope protein of the AIDS virus that may function in membrane interactions, are being actively investigated. Structural and active site experiments should help to elucidate the role of protein methyltransferase in control of eucaryotic membrane function. Also, the role of ubiquitin in the control of intracellular protein degradation in eucaryotes is being probed.